PDAS2
Introduction
DAS2 is a promoter derived from dedihydroxy-acetone synthase (DAS) gene[1].
Characterization
In order to test the function of PDAS2, we construct "PDAS2-EGFP-terminator" (Figure 1). If PDAS2 is functional, we can test the fluorescence intensity of EGFP in supernatant samples obtained from the culture of recombinant P.pastoris GS115 strain.
Figure 1 Gene circuit of PDAS2-EGFP-terminator
Our results matched the general expected trend (Figure 2). After fermentation experiment in BMMY medium containing 1% methanol. The fluorescence intensity of the samples of recombinant P.pastoris GS115 containing the EGFP gene gradually increased over time. At the same time, we measured the growth curve of the strains.
Figure 2 Fluorescence intensity and OD600 absorbance of samples obtained at different time points from the culture of corresponding recombinant P.pastoris GS115 containing EGFP gene.
Reference
[1] Vogl T, Kickenweiz T, Pitzer J, et al. Engineered bidirectional promoters enable rapid multi-gene co-expression optimization[J]. Nat Commun, 2018,9(1):3589.